This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Talarico, T L Articles by Dallas, W S Search for Related Content PubMed PubMed Citation Articles by Talarico, T L Articles by Dallas, W S

research-article

Cloning, sequence analysis, and overexpression of Escherichia coli folK, the gene coding for 7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase.

Department of Molecular Genetics and Microbiology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.

ABSTRACT

The gene coding for the Escherichia coli enzyme 7,8-dihydro-6-hydroxymethylpterin-pyrophosphokinase has been cloned and sequenced. This gene, designated folK, codes for a protein of 159 amino acids, including an amino-terminal methionine. The protein was overexpressed in E. coli MC4100 by cloning the gene behind the lacUV5 promoter in a high-copy-number plasmid. The enzyme was purified to homogeneity. Amino-terminal analysis of the purified protein showed that the amino-terminal methionine had been removed. The compositional molecular mass (17,945 Da) was identical to the molecular mass determined by mass spectrometry. The enzyme was observed to have a large number of proline residues and migrated anomalously in sodium dodecyl sulfate-polyacrylamide gels, with an apparent molecular mass of 23,000 Da.

This article has been cited by other articles:

• Guldener, U., Koehler, G. J., Haussmann, C., Bacher, A., Kricke, J., Becher, D., Hegemann, J. H. (2004). Characterization of the Saccharomyces cerevisiae Fol1 Protein: Starvation for C1 Carrier Induces Pseudohyphal Growth. Mol. Biol. Cell 15: 3811-3828 [Abstract] [Full Text]  
• Berlyn, M. K. B. (1998). Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Microbiol. Mol. Biol. Rev. 62: 814-984 [Abstract] [Full Text]  
• Bermingham, A., Bottomley, J. R., Primrose, W. U., Derrick, J. P. (2000). Equilibrium and Kinetic Studies of Substrate Binding to 6-Hydroxymethyl-7,8-dihydropterin Pyrophosphokinase from Escherichia coli. J. Biol. Chem. 275: 17962-17967 [Abstract] [Full Text]